Mycol Progress (2014) 13:771–780 DOI 10.1007/s11557-014-0960-8

ORIGINAL ARTICLE

Flammeopellis bambusicola gen. et. sp. nov. (, ) evidenced by morphological characters and phylogenetic analysis

Chang-Lin Zhao & Xin-Sheng He & Kun-Yuan Wanghe & Bao-Kai Cui & Yu-Cheng Dai

Received: 17 October 2013 /Revised: 23 December 2013 /Accepted: 9 January 2014 /Published online: 26 January 2014 # German Mycological Society and Springer-Verlag Berlin Heidelberg 2014

Abstract A new poroid wood-inhabiting fungal genus, Keywords Multi-marker analysis . . Flammeopellis, is proposed based on morphological characters . . Wood-rotting fungi and molecular evidence. The genus is typified by Flammeopellis bambusicola sp. nov., which macroscopically ischaracterizedbyanannualgrowthhabitandstipitate Introduction basidiocarps with a reddish-brown pileal cuticle. Microscopically, it has a dimitic hyphal system with generative The Polyporales is a diverse group of includ- hyphae frequently with simple septa, occasionally with clamp ing more than 1,800 described species in 216 genera and 13 connections, strongly dextrinoid and cyanophilous skeletal hy- families (Kirk et al. 2008). Polypores are a very important phae, and ellipsoid to drop-shaped, pale yellowish, thick-walled, group of wood-inhabiting fungi because of their key role in smooth, weakly dextrinoid and cyanophilous basidiospores. the carbon cycle, and the white-rot fungi of polypores are Phylogenetic analysis based on molecular data of ITS+LSU among the most efficient wood decayers of the biosphere nrRNA gene regions indicates that Flammeopellis belongs to (Zhou et al. 2011; Floudas et al. 2012). Some of these fungi the core polyporoid clade and is closely related to are also forest pathogens and have potential applications in Perenniporiella. Combined ITS+nLSU+mtSSU+TEF1 se- biomedical engineering and biodegradation (Younes et al. quence data of representative taxa in Perenniporia sensu lato 2007; Dai et al. 2007, 2009;Wangetal.2012, 2013;Cao demonstrated that Flammeopellis bambusicola grouped with et al. 2013; Si and Cui 2013;Sietal.2013). the Perenniporiella clade, but formed a monophyletic lineage Molecular studies involving have mainly been with a strong support (100 % BP, 1.00 BPP). The morphological carried out based on ITS and/or nLSU sequences (Robledo and molecular evidence confirmed the placement of the new et al. 2009; Justo and Hibbett 2011; Miettinen and Larsson genus in the core polyporoid clade and established its relation- 2011; Miettinen and Rajchenberg 2012). In addition, a six-gene ships with similar genera. dataset have helped to clarify the generic relationships of 373 polyporoid taxa and provide a phylogenetic and phylogenomic overview of the Polyporales (Binder et al. 2013). China has a huge land area, including boreal, temperate, subtropical, and tropical vegetation, and the diversity of C.

Dai 2013). During investigations on wood-inhabiting fungi in specimen was lysed in 30 μl dilution buffer for DNA extrac- southwest China, an additional undescribed was found tion. After incubating 3 min at room temperature, 0.75 μlof in Sichuan Province. It is characterized by an annual growth the supernatant were used as template for a 30-μl PCR reac- habit, stipitate basidiocarps with reddish-brown cuticle at tion. The DNAwas amplified with the primers: ITS4 and ITS5 pileal surface, a dimitic hyphal system with generative hyphae for ITS, MS1 and MS2 for mtSSU (White et al. 1990); LR0R frequently with simple septa, occasionally with clamp con- and LR7 for nLSU (http://www.biology.duke.edu/fungi/ nections, strongly dextrinoid and cyanophilous skeletal hy- mycolab/primers.htm), EF1-983 F and EF1-2218R for TEF1 phae, and ellipsoid to drop-shaped, pale yellowish, thick- (Rehner and Buckley 2005). The PCR procedure for ITS, walled, smooth, weakly dextrinoid and cyanophilous basidio- mtSSU and TEF1 were as follows: initial denaturation at spores; additionally, it grows on bamboo. These characters 98 °C for 5 min, followed by 39 cycles at 98 °C for 5 s, distinguish it from all known wood-inhabiting fungal taxa. In 58 °C for 5 s and 72 °C for 5 s, and a final extension of 72 °C this study, we expand the phylogenetic sampling of wood- for 10 min. The only difference of the nLSU amplification inhabiting poroid taxa using internal transcribed spacer (ITS) procedure was its annealing temperature was 48 °C. DNA regions and the large subunit nuclear ribosomal RNA gene sequencing was performed at Beijing Genomics Institute, (nLSU) sequences to examine taxonomy and phylogeny of China. All newly generated sequences were submitted to this new genus within the Polyporales. In addition, translation GenBank and are listed in Table 1. elongation factor 1-α (TEF1) and mitochondrial rRNA gene Sequences generated for this study were aligned with ad- sequences (mtSSU) were obtained for representative taxa in ditional sequences downloaded from GenBank (Table 1) the Perenniporia s.l., and a further investigation on the phy- using BioEdit (Hall 1999) and ClustalX (Thompson et al. logenetic relationships of the newly described genus and 1997). Sequence alignments were deposited in TreeBASE related genera was carried out. (submission ID 14809). Maximum parsimony analysis was applied to the combined multiple genes dataset sequences, and this test under heuristic Materials and methods search and 1,000 homogeneity replicates gave a P value of 1.000, much greater than 0.01, which means there is no dis- Morphological studies crepancy among the four loci in reconstructing phylogenetic trees. The tree construction procedure was performed in The studied specimens were deposited at the herbarium of PAUP* v.4.0b10 (Swofford 2002). All characters were equally Beijing Forestry University (BJFC). The microscopic exami- weighted and gaps were treated as missing data. Trees were nations followed Cui et al. (2007). Sections were studied at inferred using the heuristic search option with TBR branch magnification up to ×1,000 using a Nikon Eclipse 80i micro- swapping and 1,000 random sequence additions. Max-trees scope and phase contrast illumination (Nikon, Tokyo, Japan). were set to 5,000, branches of zero length were collapsed, Drawings were made with the aid of a drawing tube. and all parsimonious trees were saved. Clade robustness was Microscopic features, measurements, and drawings were made assessed using a bootstrap (BT) analysis with 1,000 replicates from slide preparations stained with Cotton Blue and Melzer’s (Felsenstein 1985). Descriptive tree statistics tree length (TL), reagent. Spores were measured from sections cut from the consistency index (CI), retention index (RI), rescaled consis- tubes. In presenting the variation in the size of the spores, tency index (RC), and homoplasy index (HI) were calculated 5 % of measurements were excluded from each end of the for each Maximum Parsimonious Tree (MPT) generated. range, and were given in parentheses. In the text, the following MrMODELTEST2.3 (Posada and Crandall 1998; abbreviations were used: IKI = Melzer’s reagent, KOH=5 % Nylander 2004) was used to determine the best-fit evolution potassium hydroxide, CB = Cotton Blue, CB+ = cyanophilous, model for each data set for Bayesian inference (BI). Bayesian L = mean spore length (arithmetic average of all spores), W = inference was calculated with MrBayes3.1.2 with a general mean spore width (arithmetic average of all spores), Q = vari- time reversible (GTR) model of DNA substitution and a ation in the L/W ratios between the specimens studied, n = gamma distribution rate variation across sites (Ronquist and number of spores measured from given number of specimens. Huelsenbeck 2003). Four Markov chains were run for 2 runs Special color terms follow Petersen (1996). from random starting trees for 2 million generations, and trees were sampled every 100 generations. The first one-fourth of Molecular procedures and phylogenetic analyses generations were discarded as burn-in. A majority rule con- sensus tree of all remaining trees was calculated. Branches The fungal taxa used in this study are listed in Table 1.A that received bootstrap support for maximum parsimony (MP) Phire® Plant Direct PCR Kit (Thermo, Vilnius, Lithuania) was and Bayesian posterior probabilities (BPP) greater than or used to obtain PCR products from dried specimens, according equal to 75 % (MP) and 0.95 (BPP) were considered as to the manufacturer’s instructions. About 0.5 g dried fungal significantly supported, respectively. Mycol Progress (2014) 13:771–780 773

Table 1 A list of species, specimens, and GenBank accession number of sequences used in this study

Species name Sample no. GenBank accession nos. References

ITS LSU mtSSU TEF1

Abortiporus biennis TFRI 274 EU232187 EU232235 ——Binder et al. 2005 A. biennis EL 6503-03 JN649325 JN649325 ——Sjökvist et al. 2012 Abundisporus fuscopurpureus Cui 10950 KC456254a KC456256a KF051025a KF181154a Present study A. pubertatis Cui 5776 KC787565a KC787572a KF051029a KF181129a Present study A. roseoalbus Dai 12269 KC415908a KC415910a KF051037a KF181131a Present study A. sclerosetosus MUCL 41438 FJ411101 FJ393868 ——Robledo et al. 2009 A. violaceus Ryvarden 10775 KF018126a KF018134a KF051058a KF181152a Present study Amylocystis lapponica KHL 11755 EU118603 EU118603 ——Larsson 2007 Antrodia albida CBS 308.82 DQ491414 AY515348 ——Kim et al. 2007 A. albida FP 105979 EU232272 EU232272 ——Binder et al. 2005 A. heteromorpha CBS 200.91 DQ491415 — DQ491442 Kim et al. 2007 A. macra MUAF 887 EU340898 EU340898 ——Cui 2013 Bjerkandera adusta NBRC 4983 AB733156 AB733333 ——Binder et al. 2005 Cinereomyces lindbladii FBCC 177 HQ659223 ———Miettinen and Rajchenberg 2012 Climacocystis borealis KH 13318 JQ031126 JQ031126 ——Sjökvist et al. 2012 Coriolopsis caperata LE(BIN)-0677 AB158316 AB158316 — EU03059 Justo and Hibbett 2011 Donkioporia expansa MUCL 35116 FJ411104 FJ393872 ——Robledo et al. 2009 Earliella scabrosa PR1209 JN165009 JN164793 — JN164894 Justo and Hibbett 2011 Flammeopellis bambusicola Dai 13443 KF698748a KF698759a KF725877a KF725879a Present study F. bambusicola Dai 13506 KF698749a KF698760a KF725878a KF725880a Present study Ganoderma lingzhi Wu 1006-38 JQ781858 — JX029989a JX029976a Present study G. lingzhi Dai 12479 JQ781864 — JX029988a JX029975a Present study G. australe Cui 9511 JN048773 JN048792 ——Zhao et al. 2013a G. sinense Wei 5327 KF494998a KF495008a — KF494976a Present study G. applanatum Dai 12483 KF494999a KF495009a — KF494977a Present study Gelatoporia subvermispora Kotiranta 20823 FN907911 FN907911 ——Miettinen and Larsson 2011 Grammothelopsis subtropica Cui9041 JQ845096a JQ845099a KF051039a KF181133a Present study Heterobasidion annosum PFC 5252 KC492906 KC492906 — KC571646 Binder et al. 2013 Hornodermoporus latissimus Cui 6625 HQ876604 JF706340 KF051040a KF181134a Zhao and Cui 2012 H. martius Cui 7992 HQ876603 HQ654114 KF051041a KF181135a Zhao and Cui 2012 H. martius MUCL 41677 FJ411092 FJ393859 ——Robledo et al. 2009 H. martius MUCL 41678 FJ411093 FJ393860 ——Robledo et al. 2009 Hydnopolyporus fimbriatus LR 40855 JN649347 JN649347 ——Sjökvist et al. 2012 Hypochnicium lyndoniae NL 041031 JX124704 JX124704 ——Binder et al. 2005 Lentinus tigrinus DSH93-181 AY218419 AF518627 U27050 — Wang et al. 2004 Microporellus violaceocinerascens MUCL 45229 FJ411106 FJ393874 ——Robledo et al. 2009 M. violaceocinerascens Cui 8459 HQ876606 HQ654113 KF051042a KF181136a Zhao and Cui 2013 Obba rivulosa KCTC 6892 FJ496693 FJ496710 ——Tomšovský et al. 2010 Perenniporia hainaniana Cui 6364 JQ861743 JQ861759 KF051044a KF181138a Zhao and Cui 2013 P. hainaniana Cui 6365 JQ861744 JQ861760 KF051045a KF181139a Zhao and Cui 2013 P. hainaniana Cui 6366 JQ861745 JQ861761 KF494996a KF494981a Zhao and Cui 2013 P. medulla-panis MUCL 49581 FJ411088 FJ393876 ——Robledo et al. 2009 P. medulla-panis MUCL 43250 FJ411087 FJ393875 ——Robledo et al. 2009 P. medulla-panis Cui 3274 JN112792 JN112793 KF051043a KF181137a Zhao and Cui 2012 P. subacida Cui 10053 KF495006a KF495017a KF218321a KF286327a Present study P. substraminea Dai 10781 KF495007a KF495018a KF494995a KF494983a Present study P. substraminea Cui 10177 JQ001852 JQ001844 KF051046a KF181140a Zhao and Cui 2013 774 Mycol Progress (2014) 13:771–780

Table 1 (continued)

Species name Sample no. GenBank accession nos. References

ITS LSU mtSSU TEF1

P. substraminea Cui 10191 JQ001853 JQ001845 KF051047a KF181141a Zhao and Cui 2013 Perenniporiella chaquenia MUCL 47647 FJ411083 FJ393855 — HM467609 Robledo et al. 2009 P. chaquenia MUCL 47648 FJ411084 FJ393856 — HM467610 Robledo et al. 2009 P. micropora MUCL43581 FJ411086 FJ393858 — HM467608 Robledo et al. 2009 P. neofulva MUCL 45091 FJ411080 FJ393852 — HM467599 Robledo et al. 2009 P. pendula MUCL 46034 FJ411082 FJ393853 — HM467601 Robledo et al. 2009 Phanerochaete chrysosporium BKM-F-1767 HQ188436 GQ470643 ——Binder et al. 2005 Phlebia unica KHL 11786 EU118657 EU118657 ——Larsson 2007 Physisporinus sanguinolentus BRNM 699576 FJ496671 FJ496725 FJ496750 — Tomšovský et al. 2010 Piloporia sajanensis Mannine 2733a HQ659239 HQ659239 ——Miettinen and Rajchenberg 2012 Polyporus brumalis KHL 8558 AF347108 AF347108 ——Larsson 2007 P. tuberaster CulTENN 8976 AF516598 AJ488116 ——Binder et al. 2005 Postia alni X 1400 KC595932 KC595932 ——Beatriz et al. 2013 P. caesia CIEFAP 174 JX090109 JX090130 ——Binder et al. 2013 P. guttulata KHL 11739 EU11865 EU11865 ——Beatriz et al. 2013 P. l act ea X 1391 KC595939 KC595939 ——Beatriz et al. 2013 P. venata CIEFAP 346 JX090113 JX090133 ——Beatriz et al. 2013 Pycnoporus cinnabarinus AFTOL-ID 772 DQ411525 AY586703 U27059 — Binder et al. 2005 Pyrofomes demidoffii MUCL 41034 FJ411105 FJ393873 ——Robledo et al. 2009 Sebipora aquosa Miettinen 8680 HQ659240 HQ659240 ——Miettinen and Rajchenberg 2012 Skeletocutis amorpha Miettinen 11038 FN907913 FN907913 ——Miettinen and Larsson 2011 Stereum hirsutum NBRC 6520 AB733150 AB733325 ——Binder et al. 2013 Trametes elegans FP105679 JN048766 JN048785 ——Zhao et al. 2013a T. hirsuta Cui 7784 JN048768 JN048787 ——Zhao et al. 2013a T. hirsuta RLG5133T JN164854 JN164801 AF042154 JN164891 Justo and Hibbett 2011 T. pubescens PRM 900586 AY684173 AY855906 ——Tomšovský et al. 2006 Truncospora ochroleuca Dai 11486 HQ654105 JF706349 KF051048a KF181142a Zhao and Cui 2012 T. ochroleuca MUCL 39726 FJ411098 FJ393865 ——Robledo et al. 2009 T. ochroleuca Cui 5671 JX941584 JX941602 KF218309a KF286315a Zhao and Cui 2013 T. ochroleuca Cui 5673 JX941585 JX941603 KF218308a KF286314a Zhao and Cui 2013 T. ohiensis Cui 5714 HQ654103 HQ654116 KF051056a KF181150a Zhao and Cui 2012 T. ohiensis MUCL 41036 FJ411096 FJ393863 ——Robledo et al. 2009 Tyromyces chioneus Miettinen 7487 HQ659224 HQ659224 ——Miettinen and Rajchenberg 2012 T. kmetii Penttilä 13474 KF705040a KF705041a ——Present study Vanderbylia delavayi Dai 6891 JQ861738a KF495019a KF218287a KF286293a Present study V. fraxinea DP 83 AM269789 AM269853 ——Robledo et al. 2009 V. fraxinea Cui 7154 HQ654095 HQ654110 KF218288a KF286294a Zhao and Cui 2012 V. fraxinea Cui 8871 JF706329 JF706345 KF051050a KF181144a Zhao and Cui 2012 V. fraxinea Cui 8871 JF706329 JF706345 KF051050a KF181144a Zhao and Cui 2012 V. robiniophila Cui 7144 HQ876608 JF706341 KF051052a KF181146a Zhao and Cui 2012 V. vicina MUCL 44779 FJ411095 AF518666 ——Robledo et al. 2009 Yuchengia narymica Dai 6998 JN048775 JN048794 KF051055a KF181149a Zhao et al. 2013a a Newly generated sequences for this study Mycol Progress (2014) 13:771–780 775

Results annosum (Fr.) Bref. obtained from GenBank were used as outgroups to root trees following Binder et al. (2013). Molecular phylogeny The phylogeny (Fig. 1) inferred from ITS+nLSU se- quences demonstrated six major clades for 55 sampled species Eighty-eight new gene sequences were generated in this study of the Polyporales. The new genus Flammeopellis clustered (Table 1). The ITS+nLSU dataset included sequences from 61 into the core polyporoid clade and was closely related to fungal specimens representing 55 taxa. The dataset had an Perenniporiella Decock & Ryvarden based on phylogenetic aligned length of 2,148 characters, of which 1,207 characters analysis with a low support (77 % BP, 0.97 BPP), and then are constant, 256 are variable and parsimony-uninformative, grouped with the clades of Abundisporus Ryvarden, and 685 are parsimony-informative. Maximum parsimony Ganoderma P. Karst, Grammothelopsis Jülich and analysis yielded 5 equally parsimonious trees (TL=4,520, Truncospora Pilát. CI=0.344, RI=0.517, RC=0.277, HI=0.656). Best model The four-gene (ITS+LSU+mtSSU+TEF1) sequence for ITS+nLSU estimated and applied in the Bayesian analysis: dataset did not show any conflicts in tree topology for the GTR+I+G. Bayesian analysis resulted in the same topology reciprocal bootstrap trees, which allowed us to combine them compared to maximum parsimony analysis with an average (P>0.01). The combined dataset included sequences from 50 standard deviation of split frequencies=0.007869. Sequences fungal specimens representing 31 taxa. The dataset had an of Stereum hirsutum (Willd.) Pers. and Heterobasidion aligned length of 4,035 characters, of which 2,545 characters

Fig. 1 One of the most parsimonious trees illustrating the phylogeny of Flammeopellis bambusicola and related species in Polyporales, based on ITS+ nLSU sequences. Parsimony bootstrap proportions (before the slash markers)higherthan50% and Bayesian posterior probabilities (after the slash markers) more than 0.95 are indicated along branches 776 Mycol Progress (2014) 13:771–780 are constant, 230 are variable and parsimony-uninformative, Differs from other genera by a stipitate basidiocarp with and 1,260 are parsimony-informative. Maximum parsimony reddish cuticle on pileus and stipe surface, a dimitic hyphal analysis yielded 8 equally parsimonious trees (TL=4,484, system with generative hyphae frequently with simple septa, CI=0.492, RI=0.659, RC=0.323, HI=0.509). Best model occasionally with clamp connections, strongly dextrinoid and for the combined ITS+nLSU+mtSSU+TEF1 estimated and cyanophilous skeletal hyphae, and thick-walled, pale yellowish, applied in the Bayesian analysis: GTR+I+G. Bayesian analy- smooth, weakly dextrinoid and cyanophilous basidiospores. sis resulted in the same topology with an average standard Type species: Flammeopellis bambusicola. deviation of split frequencies=0.008430. Sequences of Etymology: flammeopellis(Lat.): referring to reddish upper Donkioporia expansa (Desm.) Kotl. & Pouzar and cuticle surface. Pyrofomes demidoffii (Lév.) Kotl. & Pouzar obtained from Basidiocarps annual, stipitate, soft corky when fresh, corky GenBank were used as outgroups to root trees following when dried. Pileus convex, pileal surface bearing a reddish Zhao et al. (2013b). cuticle on pileus and stipe surface. Pore surface white to A further phylogeny (Fig. 2) inferred from multiple genes cream. Context white to cream, soft corky when fresh, corky of the combined ITS+nLSU+mtSSU+TEF1 sequences was when dried. Tubes concolorous with pore surface, soft corky obtained for representative taxa in the Perenniporia s.l. and when fresh, corky when dried. Stipe distinctly reddish, corky. demonstrated that Flammeopellis bambusicola grouped with Hyphal system dimitic; generative hyphae frequently with Perenniporiella clade, but two sampled specimens of the new simple septa, occasionally with clamp connections; skeletal species F. bambusicola, formed a monophyletic lineage with a hyphae strongly dextrinoid, CB+. Basidiospores pale yellow- strong support (100 % BP, 1.00 BPP), and then grouped with ish, thick-walled, smooth, weakly dextrinoid, CB+. Grammothelopsis subtropica B.K. Cui & C.L. Zhao with a Flammeopellis bambusicola Y.C. Dai, B.K. Cui & C.L. 94 % bootstrap value and 0.99 Bayesian posterior probability. Zhao, sp. nov. Figs. 3 and 4 MycoBank no.: MB 807158 Basidiocarps annual, stipitate, solitary or gregarious, soft Taxonomy corky when fresh, corky when dried. Pilei more or less semi- circular to spathulate. Pileus convex, pileal surface bearing a Flammeopellis Y.C. Dai, B.K. Cui & C.L. Zhao, gen. nov. reddish brown cuticle. Pore surface white when fresh, cream MycoBank no.: MB 807157 upon drying; pores round; dissepiments thick, entire. Context

Fig. 2 One of the most parsimonious trees illustrating the phylogeny of Flammeopellis bambusicola species and related species obtained for more representative taxa in the Perenniporia s.l., based on the combined ITS+LSU+mtSSU+ TEF1 sequence datasets. Parsimony bootstrap proportions (before the slash markers)higher than 50 % and Bayesian posterior probabilities (after the slash markers) more than 0.95 are indicated along branches Mycol Progress (2014) 13:771–780 777

yellowish, thick-walled, smooth, weakly dextrinoid, cyanophilous, 4.5–5.1×3.5–4 μm. Holotype: CHINA. Sichuan Prov., Qionglai County, Pingle, Lugouzhuhai, on stump of Bambusa, 12 August 2013 Dai 13443 (BJFC). Etymology: bambusicola (Lat.): referring to growth on bamboo stump. Basidiome: Basidiocarps annual, laterally stipitate, solitary or gregarious, soft corky and without odor or taste when fresh, becoming corky upon drying. Pilei more or less semicircular to spathulate, up to 7 cm long, 5 cm wide, 4 mm thick at centre. Pileus convex, pileal surface bearing a reddish brown cuticle which becoming dark reddish brown with age, irregu- larly rough, distinctly sulcate. Pore surface white when fresh, Fig. 3 Basidiocarps of Flammeopellis bambusicola (holotype). Bars (a) white to cream upon drying; pores round, 6–7 per mm; dis- 1.5 cm, (b)0.5cm,(c)2cm sepiments thick, entire. Sterile margin narrow, cream to pale brown, up to 1 mm wide. Context white to cream, soft corky when fresh, corky when dried, up to 1.5 mm thick. Tubes white to cream, soft corky when fresh, corky when dried. concolorous with pore surface, soft corky when fresh, corky Tubes concolorous with pore surface, soft corky when fresh, when dried, up to 2.5 mm long. Stipe surface distinctly red- corky when dried. Stipe surface distinctly reddish, corky. dish, soft corky when fresh, corky when dried, up to 4.5 cm Tissues unchanged in KOH. Hyphal system dimitic; genera- long, 1.5 mm in diam. tive hyphae frequently with simple septa, occasionally with Hyphal structure: Hyphal system dimitic; generative hy- clamp connections; skeletal hyphae strongly dextrinoid, CB+, phae frequently with simple septa, occasionally with clamp unbranched. Basidiospores ellipsoid to drop-shaped, pale connections; skeletal hyphae strongly dextrinoid, CB+; tissues unchanged in KOH. Context: Generative hyphae infrequent, hyaline, thin-walled, occasionally branched, 3.5–5 μm in diameter; skeletal hyphae dominant, hyaline, thick-walled with a wide lumen, unbranched, interwoven, occasionally collapsed, 4–6 μm in diameter. Cuticle: Composed of a vertical and closely-packed palisade of cells; cells mostly clavate, yellowish to pale brown, thick- walled, with 1–2 septa, weakly dextrinoid, 31–45×5–7 μm. Tubes: Generative hyphae infrequent, hyaline, thin-walled, occasionally branched, 3–4.5 μm in diameter; skeletal hyphae dominant, hyaline, thick-walled with a wide to narrow lumen, unbranched, interwoven, 3.5–5 μm in diameter. Cystidia absent, but fusoid cystidioles present, hyaline, thin-walled, 10–14×3– 4 μm; basidia clavate to barrel-shaped, with four sterigmata and a basal clamp connection, 15–16×5–6.5 μm; basidioles domi- nant, in shape similar to basidia, but slightly smaller. Stipe: Generative hyphae infrequent, hyaline, thin-walled, occasionally branched, 3–5 μm in diameter; skeletal hyphae dominant, hyaline, thick-walled with a wide lumen, un- branched, interwoven, 3.5–6 μmindiameter. Spores: Basidiospores ellipsoid to drop-shaped, pale yellowish, thick-walled, smooth, weakly dextrinoid, CB+, (4.3–)4.5–5.1(−5.4)×(3.3–)3.5–4(−4.2) μm, L =4.83μm, W =3.8μm, Q =1.25–1.28 (n =60/2). Rot type: White rot. Fig. 4 Microscopic structures of Flammeopellis bambusicola (drawn Additional specimen examined: CHINA. Sichuan Prov., from the holotype). a Basidiospores; b basidia and basidioles; c cystidioles; d palisade of cells from the upper surface cuticle; e hyphae Qionglai County, Pingle, Lugouzhuhai, on stump of Bambusa, from trama; f hyphae from context. Bars:(a)5μm, (b–f)10μm 10 September 2013 Dai 13506 (BJFC, paratype). 778 Mycol Progress (2014) 13:771–780

Discussion bambusicola differs in its resupinate basidiocarps with orange to orange brown pore surface, presence of rhizomorphs, and Flammeopellis bambusicola is characterized by an annual truncate, hyaline, narrower basidiospores (1.8–2.5 μmwide; growth habit, laterally stipitate basidiocarps with a reddish- Choeyklin et al. 2009). Perenniporia aridula is distinguished brown pileal cuticle, a dimitic hyphal system with generative by perennial, resupinate basidiocarps, frequently branched hyphae frequently with simple septa, occasionally with clamp skeletal hyphae, and truncate and larger basidiospores (6–7× connections, strongly dextrinoid, cyanophilous, unbranched 5.1–6 μm; Zhao et al. 2013b). skeletal hyphae, and ellipsoid to drop-shaped, pale yellowish, thick-walled, smooth, weakly dextrinoid and cyanophilous Acknowledgments The research is supported by the Fundamental basidiospores (4.5–5.1×3.5–4 μm). Research Funds for the Central Universities (Project No. JC2013-1), Beijing In the present study, Flammeopellis bambusicolais described Higher Education Young Elite Teacher Project (YETP0774) and the National Natural Science Foundation of China (Project No. 31093440). based on morphological characters and molecular phylogenetic analyses. The molecular data show that Flammeopellis belongs to the core polyporoid clade and is closely related to Perenniporiella based on ITS+LSU-nrRNA gene regions with References a low support (77 % BP, 0.97 B.P. Fig. 1). A further study with representative taxa in the Perenniporia s.l. used combined ITS+ Beatriz OS, Lindner DL, Miettinen O, Justo A, Hibbett DS (2013) A nLSU+mtSSU+TEF1 sequence data analysis. Flammeopellis phylogenetic overview of the antrodia clade (Basidiomycota, Polyporales). Mycologia 105:1391–1411. doi:10.3852/13-051 bambusicola grouped with the Perenniporiella clade, but Binder M, Hibbett DS, Larsson KH, Larsson E, Langer E, Langer G formed a monophyletic lineage with a strong support (100 % (2005) The phylogenetic distribution of resupinate forms across the BP, 1.00 B.P. Fig. 2). Flammeopellis is therefore confirmed as a major clades of mushroom-forming fungi (Homobasidiomycetes). – new genus in the core polyporoid clade. Syst Biodivers 3:113 157. doi:10.1017/S1477200005001623 Binder M, Justo A, Riley R, Salamov A, López-Giráldez F, Sjökvist E, Phylogenetically, the genus Perenniporiella groups with Copeland A, Foster B, Sun H, Larsson E, Larsson KH, Townsend J, Flammeopellis (Figs. 1, 2); however, the former differs from Grigoriev IV, Hibbett DS (2013) Phylogenetic and phylogenomic the latter by its dimitic hyphal system, generative hyphae with overview of the Polyporales. 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